Method for enhancing gene expression
AssigneeGenentech, Inc.
InventorFlorian M. Wurm
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Abstract
Methods are provided for the identification and use of endogenous nucleic acid sequences for homologous recombination and targeted integration of heterologous nucleic acid sequences. These methods are useful for introduction of nucleic acid into cells or tissue for gene therapy, or for the preparation of transgenic animals. Methods are also provided for enhancing the yield of genes of interest from recombinant cell culture lines.
Claims14
A method for obtaining animal cells having increased expression of a heterologous gene comprising:
- a. obtaining an animal cell having a first endogenous nucleic acid sequence which is present in at least 50 copies and dispersed within the genome of the host cell and is present in transcriptionally active areas of the genome;
- and
- b. introducing into the cell a second nucleic acid sequence homologous to the endogenous sequence and a third nucleic acid sequence which encodes a heterologous gene.
A method for obtaining animal cells having increased expression of a heterologous gene comprising:
- a. identifying a first nucleic acid sequence which is endogenous in an animal cell and which is present in at least 50 copies dispersed through the genome of the cell and is present in transcriptionally active areas of the genome;
- and
- b. introducing into the cell a second nucleic acid sequence homologous to the first nucleic acid sequence and a third nucleic acid sequence which encodes a heterologous gene.
A method for obtaining animal cells having increased expression of a heterologous gene comprising:
- a. obtaining an animal cell having a first endogenous nucleic acid sequence which is present in at least 50 copies in the animal cell genome, the copies being present in transcriptionally active regions of the genome, wherein any two copies are separated from each other by being located on different chromosomes or located at least 100,000 base pairs apart on the same chromosome;
- and
- b. introducing into the cell a second nucleic acid sequence homologous to the endogenous sequence and a third nucleic acid sequence which encodes a heterologous gene.
Dependencies are inferred from the claim text.